ANALYSIS OF 4-HYDROXYCYCLOPHOSPHAMIDE IN CANCER PATIENTS PLASMA FOR THERAPEUTIC DRUG MONITORING OF CYCLOPHOSPHAMIDE
Objective: To quantify 4-hydroxycyclophosphamide in cancer patients' plasma for therapeutic drug monitoring of cyclophosphamide.
Methods: The blood was collected at 0.5 and 1 h after administration of chemotherapy. Prior to analysis, 4-OHCP in plasma was derivatized with semicarbazide HCl, then was extracted using 4 ml ethyl acetate and finally was determined by Ultra High-Performance Liquid Chromatographyâ€“tandem mass spectrometry. Chromatographic separation was conducted using waters acquity BEH C18 column (1.7 Î¼m; 50 mm x 2.1 mm). The mobile phase consisted of formic acid 0.1% and methanol (50: 50, v/v), column temperature 30 Â°C and flow rate of 0.3 ml/min. Mass detection was performed on waters xevo TQD equipped with an electrospray ionization (ESI) source at positive ion mode in the multiple reaction monitoring (MRM). Cyclophosphamide was detected at m/z 260.968>139.978, 4-hydroxycyclophosphamide-semicarbazide at m/z 338.011>224.97, and hexamethylphosphoramide as internal standard at m/z 180.17>92.08.
Results: The method was linear in the range of 5â€“1000 ng/ml for cyclophosphamide and also for 4-hydroxycyclophosphamide. The results showed that the level of 4-OHCP in 39 cancer patients was in the range of 5.02 ng/ml to 832.44 ng/ml.
Conclusion: 4-hydroxycyclophosphamide was detected on 39 patient samples with the lowest level of 5.40ng/ml and the highest level was 832.44 ng/ml. This can be a parameter that the regiment of cyclophosphamide was effective.
2. World Health Organization. World Cancer Report. Lyon: International Agency for Research on Cancer; 2008.
3. Kementrian Kesehatan Republik Indonesia. Profil Kesehatan Indonesia Tahun. Jakarta; 2013.
4. AvendanÃµ C, MenÃ©ndez JC. Medicinal chemistry of anticancer drugs. Oxford: Elsevier; 2008.
5. De Jonge ME, Huitema AD, Rodenhuis S, Beijnen JH. Clinical pharmacokinetics of cyclophosphamide. Clin Pharmacokinet 2005;44:1135â€“64.
6. Nakajima Miki. Genetic polymorphisms of CYP2B6 affect the pharmacokinetics/pharmacodynamics of cyclophosphamide in Japanese cancer patients. Pharmacogenet Genomics 2007;17: 431â€“45.
7. Raccor Brianne S, Adam J Claessens, Jean C Dinh, Julie R Park, Douglas S Hawkins, Sushma S Thomas, et al. Potential contribution of cytochrome P450 2B6 to Hepatioc 4-Hydorxycyclophosphamide formation in vitro and in vivo. Drug Metab Dispos 2012;40:54â€“63.
8. Shimada T, Yamazaki H, Mimura M, Inui Y, Guengerich FP. Interindividual variations in human liver cytochrome P450 enzymes involved in the oxidation of drugs, carcinogens, and toxic chemicals: studies with liver microsomes of 30 Japanese and 30 Caucasions. J Pharmacol Exp Ther 1994;270:414-23.
9. Joy Melanie S, La Mary, Wang Jinzhao, Bridges Arlene S, Hu Yichun, Hogan Susan L, et al. Cyclophosphamide and 4-hydroxycyclophosphamide pharmacokinetics in patients with glomerulonephritis secondary to lupus and small vessel vasculitis. Br J Clin Pharmacol 2012;74:445â€“55.
10. Ekhart Corine, Gebretensae Abadi, Rosing Hilde, Rodenhuis Sjoerd, Beijnen Jos H, Huitema Alwin DR. Simultaneous quantification of cyclophosphamide and its active metabolite 4-hydroxycyclophosphamide in human plasma by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LCâ€“SM/SM). J Chromatogr B 2007;854:345â€“9.
11. Shu Wenying, Wang Xueding, Yang Xiuyan, Liang Liuqin, Li Jiali, Chen Zhuo Jia, et al. Simultaneous determination of cyclophosphamide and 4-hydroxycyclophosphamide in human plasma by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometryâ€“application to chinese systemic lupus erythematosus patients. Clin Chem Lab Med 2011;49:2029â€“37.
12. European Medicines Agency: Committee for Medicinal Products for Human Use (CHMP). Guideline on bioanalytical method validation. London: European Medicines Agency; 2012.
13. Struck Robert F, Alberts David S, Horne Katherine, Phillips J Gregory, Peng Yei-Mei, Roe Denise J. Plasma pharmacokinetics of cyclophosphamide and its cytotoxic metabolites after intravenous vs oral administration in a randomized. Crossover Trial Cancer Res 1987;47:2723-6.